The Affibody ligand also inhibits the 3‡→5‡ exonuclease activity of the polymerase, preventing degradation of primers and template DNA during reaction set up. At polymerization temperatures, the ligand is released, rendering the polymerase fully active. Phusion Hot Start II DNA Polymerase does not require a separate activation step in the PCR protocol as it is immediately reactivated at high temperatures.

New Phusion Green Hot Start II High-Fidelity DNA Polymerase is a combination of Phusion Hot Start II DNA Polymerase and 5X Phusion Green Buffers. The buffers include a density reagent and two tracking dyes for direct loading of PCR products on a gel. The colored buffer does not interfere with excelent performance of Phusion Hot Start II DNA Polymerase and is compatible with downstream applications such as DNA sequencing, ligation and restriction digestion.

Thermo Scientific Phusion Hot Start II High Fidelity DNA Polymerase is the most accurate hot start DNA polymerase on the market. It combines the Phusion DNA Polymerase and a reversibly bound, specific Affibody ligand. Affibody inhibits the DNA polymerase activity at room temperature and thus prevents the amplification of nonspecific products. The Affibody ligand also inhibits the 3'→5' exonuclease activity of the polymerase, preventing degradation of primers and template DNA during reaction set up. At polymerization temperatures, the ligand is released, rendering the polymerase fully active. Phusion Hot Start II DNA Polymerase does not require a separate activation step in the PCR protocol as it is immediately reactivated at high temperatures.New Phusion Green Hot Start II High-Fidelity DNA Polymerase is a combination of Phusion Hot Start II DNA Polymerase and 5X Phusion Green Buffers. The buffers include a density reagent and two tracking dyes for direct loading of PCR products on a gel. The colored buffer does not interfere with excelent performance of Phusion Hot Start II DNA Polymerase and is compatible with downstream applications such as DNA sequencing, ligation and restriction digestion.

Highlights

Reaction set up can be done at room temperature

Reduces non-specific amplification

Prevents primer degradation during set up

Zero-time reactivation due to unique hot start technology

Extreme fidelity (52X Taq)

Speed of the polymerase allows short extension times

Robust reactions, minimal optimization needed

Increased product yields with minimal enzyme amounts

Direct loading on gel with Green Buffer

Applications

High-fidelity PCR

High throughput

Difficult (GC-rich) templates

Template generation for sequencing

Multiplex PCR

Long-range PCR

Cloning

Mutagenesis†

Microarray

†Phusion Site-Directed Mutagenesis Kit also available.

IncludesPhusion Hot Start II High-Fidelity DNA Polymerase

Phusion Hot Start II High-Fidelity DNA Polymerase (2 U/μL)

5X Phusion HF Buffer

5X Phusion GC Buffer

DMSO

50 mM MgCl2 solution

Phusion Green Hot Start II High-Fidelity DNA Polymerase

Phusion Hot Start II High-Fidelity DNA Polymerase (2 U/μL)

5X Phusion Green HF Buffer

5X Phusion Green GC Buffer

DMSO

50 mM MgCl2 solution

Both Phusion and Phusion Green Buffers provide 1.5 mM MgCl2 in the final 1X concentration.